Process for preparing a dry powder formulation comprising an anticholinergic, a corticosteroid and a beta-adrenergic

ABSTRACT

Dry powder formulations for inhalation containing a combination of an anticholinergic, a long-acting beta2-adrenoceptor agonist, and a corticosteroid are useful for the prevention and/or treatment of an inflammatory and/or obstructive airways disease.

CROSS REFERENCES TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.15/351,562, filed on to Nov. 15, 2016, and claims priority to EuropeanPatent Application No. 15194660.5, filed on Nov. 16, 2015, which isincorporated herein by reference in its entirety.

BACKGROUND OF THE INVENTION Field of the Invention

The present invention relates to powder formulations for administrationby inhalation by means of a dry powder inhaler. In particular, thepresent invention relates to processes for preparing a dry powderformulation comprising a combination of an anticholinergic, abeta-adrenoceptor agonist, and an inhaled corticosteroid.

Discussion of the Background

Respiratory diseases are a common and important cause of illness anddeath around the world. In fact, many people are affected byinflammatory and/or obstructive lung diseases, a category characterizedby inflamed and easily collapsible airways, obstruction to airflow,problems exhaling and frequent medical clinic visits andhospitalizations. Types of inflammatory and/or obstructive lung diseaseinclude asthma, bronchiectasis, bronchitis and chronic obstructivepulmonary disease (COPD).

In particular, chronic obstructive pulmonary disease (COPD) is amulti-component disease characterized by airflow limitation and airwayinflammation. Exacerbations of COPD have a considerable impact on thequality of life, daily activities and general well-being of patients andare a great burden on the health system. Thus, the aim of COPDmanagement includes not only relieving symptoms and preventing diseaseprogression but also preventing and treating exacerbations.

While available therapies improve clinical symptoms and decrease airwayinflammation, they do not unequivocally slow long-term progression oraddress all disease components. With the burden of COPD continuing toincrease, research into new and improved treatment strategies tooptimize pharmacotherapy is ongoing, and in particular, combinationtherapies, with a view to their complementary modes of action enablingmultiple components of the disease to be addressed. Evidence from recentclinical trials indicates that triple therapy, combining ananticholinergic with an inhaled corticosteroid, and a long-actingβ₂-adrenoceptor agonist, may provide clinical benefits additional tothose associated with each treatment alone in patients with more severeCOPD.

Currently, there are several recommended classes of therapy for COPD, ofwhich bronchodilators such as β-agonists and anticholinergics are themainstay of symptom management in mild and moderate diseases, prescribedon an as-needed basis for mild COPD and as a maintenance therapy formoderate COPD.

Said bronchodilators are efficiently administered by inhalation, thusincreasing the therapeutic index and reducing side effects of the activematerial.

For the treatment of more severe COPD, guidelines recommend the additionof inhaled corticosteroids (ICSs) to long-acting bronchodilator therapy.Combinations of therapies have been investigated with a view to theircomplementary modes of action enabling multiple components of thedisease to be addressed. Data from recent clinical trials indicates thattriple therapy, combining an anticholinergic with a long-actingβ₂-agonist (LABA), and an ICS, may provide clinical benefits additionalto those associated with each treatment alone in patients with moderateto severe forms of respiratory diseases, particular moderate to severeCOPD.

An interesting triple combination, presently under investigation,includes:

-   -   i) formoterol, particularly its fumarate salt (hereinafter        indicated as FF), a long acting beta-2 adrenergic receptor        agonist, currently used clinically in the treatment of asthma,        COPD and related disorders;    -   ii) glycopyrronium bromide, an anticholinergic (antimuscarinic)        recently approved for the maintenance treatment of COPD; and    -   iii) beclometasone dipropionate (BDP) a potent anti-inflammatory        corticosteroid, available under a wide number of brands for the        prophylaxis and/or treatment of asthma and other respiratory        disorders.

The solution formulation for administration by pressurized metered doseinhalers (pMDI) is disclosed in WO 2011/076843, which is incorporatedherein by reference in its entirety.

Said formulation provides a high lung deposition and uniformdistribution throughout the bronchial tree, and is characterized by thefact that is capable of delivering a high fraction of particles having adiameter equal or less than 2.0 micron for all the three activeingredients (hereinafter defined as extrafine fraction).

The major advantage of said formulation is related to the improvedpenetration into the bronchiole-alveolar distal part of the respiratorytree wherein inflammation is known to play a role in spontaneousexacerbations of asthma symptoms and wherein it is known that thedensity of the beta-2 adrenergic receptors is particularly high.

However, despite their popularity, pMDI formulations may have somedisadvantages in particular in elderly and pediatric patients, mostlydue to their difficulty to synchronize actuation from the device withinspiration.

Dry powder inhalers (DPIs) constitute a valid alternative to MDIs forthe administration of drugs to airways.

On the other hand, drugs intended for inhalation as dry powders shouldbe used in the form of micronized particles. Their volumetriccontribution could represent an obstacle to the design of a formulationtherapeutically equivalent to one wherein the drugs are delivered informof liquid droplets.

Powder formulations for inhalation containing all said three activeingredients in a fixed combination are disclosed in WO 2015/004243,which is incorporated herein by reference in its entirety. Saidformulation takes advantage of the technology platform disclosed in WO01/78693, which is incorporated herein by reference in its entirety,entailing the use of carrier constituted of a fraction of coarseexcipient particles and a fraction made of fine excipient particles andmagnesium stearate.

In particular the teaching of WO 2015/044243, which is incorporatedherein by reference in its entirety, is mainly focused at providing an“extrafine” powder formulation wherein all the active ingredients havevery small particle size to deeply reach the distal tract of therespiratory tree.

On the other hand, the aforementioned formulation has been tailored foradministration with NEXThaler, a dry powder inhaler specificallydesigned to generate extrafine particles, and hence being particularlyefficient (see Corradi M et al Expert Opin Drug Deliv 2014, 11(9),1497-1506, which is incorporated herein by reference in its entirety).

Accordingly, the formulation of WO 2015/004243, which is incorporatedherein by reference in its entirety, loaded in highly performing drypowder inhaler may turn out to be too efficient to match theperformances of the corresponding pMDI formulation in form of solution,and hence its therapeutic characteristics.

Thus, there remains a need for powder formulations suitable for highlyperforming dry powder inhalers (DPIs) comprising formoterol fumarate,glycopyrronium bromide, and BDP in combination, overcoming the problemsindicated above and in particular to provide a powder formulation havingtherapeutic characteristics matching those of the corresponding pMDIformulation in form of solution.

SUMMARY OF THE INVENTION

Accordingly, it is one object of the present invention to provide novelpowder formulations suitable for highly performing dry powder inhalers(DPIs) comprising formoterol fumarate, glycopyrronium bromide, and BDPin combination.

It is another object of the present invention to provide novel powderformulations suitable for highly performing dry powder inhalers (DPIs)comprising formoterol fumarate, glycopyrronium bromide, and BDP incombination, overcoming the problems indicated above and in particularto provide a powder formulation having therapeutic characteristicsmatching those of the corresponding pMDI formulation in form ofsolution.

It is another object of the present invention to provide novel methodsof preparing such a formulation.

It is another object of the present invention to provide novel methodsof preventing and/or treating a disease of the respiratory tract byadministering such a formulation.

These and other objects, which will become apparent during the followingdetailed description, have been achieved by the inventors' discovery ofa process for preparing a powder formulation for inhalation for use in adry powder inhaler, said powder comprising:

(A) a carrier, comprising:

(a) 80 to 95 percent by weight, based on the total weight of saidcarrier, of coarse particles of a physiologically acceptable excipienthaving a mean particle size of at least 175 μm; and

(b) 19.6 to 4.9 percent by weight, based on the total weight of saidcarrier, of micronized particles of a physiologically acceptableexcipient, and 0.1 to 0.4 percent by weight, based on the total weightof said carrier, of a salt of a fatty acid; and

(B) micronized particles of an anti-muscarinic drug, a long-actingβ₂-agonist (LABA), and optionally, an inhaled corticosteroid (ICS), asactive ingredients,

said process comprising:

(i) mixing all of said coarse particles of a physiologically acceptableexcipient, all of said salt of a fatty acid, a first portion of saidmicronized particles of a physiologically acceptable excipient, all ofsaid micronized particles of said long-acting β₂-agonist, saidanti-muscarinic drug, and, optionally, said inhaled corticosteroid in avessel of a shaker mixer at a speed of rotation not lower than 16 r.p.m.for a time of not less than 60 minutes, to obtain a first mixture; and

(ii) adding the remaining part of said micronized particles of aphysiologically acceptable excipient to said first mixture, to obtain asecond mixture, and mixing said second mixture at a speed of rotationnot lower than 16 rpm for a time of at least 120 minutes.

In a preferred embodiment, the anti-muscarinic drug is glycopyrroniumbromide, the ICS is beclometasone dipropionate, the LABA is formoterolfumarate dihydrate, and the additive is magnesium stearate.

Therefore, in a second aspect, the present invention is directed to apowder formulation for inhalation for use in a dry powder inhaler, saidpowder comprising:

(A) a carrier, comprising:

(a) 80 to 95 percent by weight, based on the total weight of saidcarrier, of coarse particles of a physiologically acceptable excipienthaving a mean particle size of at least 175 μm; and

(b) 19.6 to 4.9 percent by weight, based on the total weight of saidcarrier, of micronized particles of a physiologically acceptableexcipient, and 0.1 to 0.4 percent by weight, based on the total weightof said carrier, of magnesium stearate; and

(B) micronized particles of glycopyrronium bromide, beclometasonedipropionate, and formoterol fumarate dihydrate, as active ingredients,

wherein said formulation is obtainable by a process comprising:

(i) mixing all of said coarse particles of a physiologically acceptableexcipient, all of said magnesium stearate, a first portion of saidmicronized particles of a physiologically acceptable excipient, all ofsaid micronized particles of glycopyrronium bromide, beclometasonedipropionate, and formoterol fumarate dihydrate in a vessel of a shakermixer at a speed of rotation not lower than 16 rpm for a time of notless than 60 minutes, to obtain a first mixture; and

(ii) adding the remaining part of said micronized particles of aphysiologically acceptable excipient to said first mixture, to obtain asecond mixture, and mixing said second mixture at a speed of rotationnot lower than 16 for a time of at least 120 minutes; whereby theextrafine particle fraction of each active ingredient is comprisedbetween 20 and 35%.

In a third aspect, the present invention concerns a dry powder inhalerdevice filled with the above dry powder formulations. Preferably the drypowder inhaler is a high-performing dry powder inhaler.

In a fourth aspect, the present invention refers to the claimedformulations for use in the prevention and/or treatment of aninflammatory and/or obstructive airways disease, in particular asthma orchronic obstructive pulmonary disease (COPD).

In a fifth aspect, the present invention provides a method for theprevention and/or treatment of an inflammatory and/or obstructiveairways disease, in particular asthma or chronic obstructive pulmonarydisease (COPD), comprising administering by inhalation, to a subject inneed thereof, an effective amount of the formulations of the presentinvention.

In a sixth aspect, the present invention refers to the use of theclaimed formulations in the manufacture of a medicament for theprevention and/or treatment of an inflammatory and/or obstructiveairways disease, in particular asthma or chronic obstructive pulmonarydisease (COPD).

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

As used herein, the term “dry powder inhaler (DPI)” refers to a devicethat delivers medication to the lungs in the form of a dry powder DPIscan be divided into two basic types:

i) single dose inhalers, for the administration of pre-subdivided singledoses of the active compound;

ii) multidose dry powder inhalers (MDPIs), either with pre-subdividedsingle doses or pre-loaded with quantities of active ingredientsufficient for multiple doses; each dose is created by a metering unitwithin the inhaler.

On the basis of the required inspiratory flow rates (1/min) which inturn are strictly depending on their design and mechanical features,DPI's are also divided in:

i) low-resistance devices (>90 l/min);

ii) medium-resistance devices (about 60-90 l/min);

iii) medium-high resistance devices (about 50-60 l/min);

iv) high-resistance devices (less than 30 l/min).

The reported classification is generated with respect to the flow ratesrequired to produce a pressure drop of 4 KPa (KiloPascal) in accordanceto the European Pharmacopoeia (Eur Ph), which is incorporated herein byreference in its entirety.

As used herein, the term “high-performing dry powder inhaler (DPI)”refers to a medium or high resistance breath-actuated multidose drypowder inhaler having a body with a mouthpiece and provided with adeagglomerator system for deagglomerating the powdered medicamentcomprising a vortex chamber (cyclone), wherein the air flow for thedelivery of the medicament is not lower than 20 l/min, preferably in therange 25 to 40 l/min.

The term is “muscarinic receptor antagonists”, “anti-muscarinic drugs”and “anticholinergic drugs” can be used synonymously.

The term “pharmaceutically acceptable salt of glycopyrrolate” refers toa salt of the compound (3S,2′R),(3R,2'S)-3-[(cyclopentylhydroxyphenylacetyl)oxy]-1,1-dimethylpyrrolidiniumin approximately 1:1 racemic mixture, also known as glycopyrronium salt.

The term “pharmaceutically acceptable salt of formoterol” refers to asalt of the compound 2′-hydroxy-5′-5-[(RS)-1-hydroxy-2{[(RS)-p-methoxy-α-methylphenethyl]amino}ethyl] formanilide.

The term “beclometasone dipropionate” refers to the compound(8S,9R,10S,11S,13S,14S,16S,17R)-9-chloro-11-hydroxy-10,13,16-trimethyl-3-oxo-17-[2-(propionyloxy)acetyl]-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[α]phenanthren-17-ylpropionate.

The term “pharmaceutically acceptable salt” comprises inorganic andorganic salts. Examples of organic salts may include formate, acetate,trifluoroacetate, propionate, butyrate, lactate, citrate, tartrate,malate, maleate, succinate, methanesulfonate, benzenesulfonate,xinafoate, pamoate, and benzoate. Examples of inorganic salts mayinclude fluoride chloride, bromide, iodide, phosphate, nitrate andsulphate.

The term “physiologically acceptable excipient” refers to apharmacologically-inert substance to be used as a carrier. In thecontext of the present invention, salts of fatty acids, that are alsophysiologically acceptable excipients, are defined as an additive.

The expression “shaker mixer” refers to a versatile mixer having a wideand adjustable range of speed of rotation and inversion cycles. In saidmixers, the mixing container is gimbal-mounted. Two rotation axes arepositioned perpendicularly each other, and are powered independently.The turning direction and rotational speed of both axes is subject tocontinual and independent change. The setting of these kind of mixingprocess parameters is able to guarantee an high value of mixingefficiency. A typical shaker mixer is commercially available asdyna-MIX™ (Willy A. Bachofen A G, Switzerland) or 3D.S mixer (ErhardMuhr GmbH, Germany).

The expression “tumbler mixer” refers to a mixer that works withdifferent mixing times and mixing speeds and but with a typical movementcharacterized by the interaction of rotation, translation and inversion.A typical tumbler mixer is commercially available as Turbula™ (Willy ABachofen AG. Switzerland).

The expression instant or high-shear mixer refers to mixers wherein arotor or impeller, together with a stationary component known as astator is used either in a tank containing the powder to be mixed tocreate a shear.

Typical high-shear mixers are P 100 and P 300 (Diosna GmbH, Germany),Roto Mix (IMA, Italy), and Cyclomix™ (Hosokawa Micron Group Ltd, Japan).

The term “micronized” refers to a substance having a size of fewmicrons.

The term “coarse” refers to a substance having a size of one or fewhundred microns.

In general terms, the particle size of particles is quantified bymeasuring a characteristic equivalent sphere diameter, known as volumediameter, by laser diffraction.

The particle size can also be quantified by measuring the mass diameterby means of suitable known instrument such as, for instance, the sieveanalyzer.

The volume diameter (VD) is related to the mass diameter (MD) by thedensity of the particles (assuming a size independent density for theparticles).

In the present application, the particle size of the active ingredientsand of fraction of fine particles is expressed in terms of volumediameter, while that of the coarse particles is expressed in terms ofmass diameter.

The particles have a normal (Gaussian) distribution which is defined interms of the volume or mass median diameter (VMD or MMD) whichcorresponds to the volume or mass diameter of 50 percent by weight ofthe particles, and, optionally, in terms of volume or mass diameter of10% and 90% of the particles, respectively.

Another common approach to define the particle size distribution is tocite three values:

i) the median diameter d(0.5) which is the diameter where 50% of thedistribution is above and 50% is below;

ii) d(0.9), where 90% of the distribution is below this value; and

iii) d(0.1), where 10% of the distribution is below this value.

The span is the width of the distribution based on the 10%, 50% and 90%quantile and is calculated according to the formula.

${Span} = \frac{{D\left\lbrack {v,0.9} \right\rbrack} - {D\left\lbrack {v,0.1} \right\rbrack}}{D\left\lbrack {v,0.5} \right\rbrack}$

In general terms, particles having the same or a similar VMD or MMD canhave a different particle size distribution, and in particular adifferent width of the Gaussian distribution as represented by thed(0.1) and d(0.9) values.

Upon aerosolization, the particle size is expressed as mass aerodynamicdiameter (MAD), while the particle size distribution is expressed interms of Mass median aerodynamic diameter (MMAD) and Geometric StandardDeviation (GSD). The MAD indicates the capability of the particles ofbeing transported suspended in an air stream. The MMAD corresponds tothe mass aerodynamic diameter of 50 percent by weight of the particles.

In the final formulation the particle size of the active ingredients canbe determined by scanning electron microscopy according to methods knownto the skilled person in the art.

The term “hard pellets” refers to spherical or semispherical units whosecore is made of coarse excipient particles.

The term “spheronization” refers to the process of rounding off of theparticles which occurs during the treatment.

The term “good flowability” refers to a formulation that is easy handledduring the manufacturing process and is able of ensuring an accurate andreproducible delivery of the therapeutically effective dose.

Flow characteristics can be evaluated by different tests such as angleof repose, Carr's index, Hausner ratio or flow rate through an orifice.

In the context of the present application the flow properties weretested by measuring the flow rate through an orifice according to themethod described in the European Pharmacopeia (Eur. Ph.) 8.6, 8^(th)Edition, which is incorporated herein by reference in its entirety.

The expression “good homogeneity” refers to a powder wherein, uponmixing, the uniformity of distribution of a component, expressed ascoefficient of variation (CV) also known as relative standard deviation(RSD), is less than 5.0%. It is usually determined according to knownmethods, for instance by taking samples from different parts of thepowder and testing the component by HPLC or other equivalent analyticalmethods.

The expression “respirable fraction” refers to an index of thepercentage of active particles which would reach the lungs in a patient.

The respirable fraction is evaluated using a suitable in viro apparatussuch as Andersen Cascade Impactor (ACI), Multi Stage Liquid Impinger(MLSI) or Next Generation Impactor (NGI), according to proceduresreported in common Pharmacopoeias, in particular in the EuropeanPharmacopeia (Eur. Ph.) 8.4, 8^(th) Edition, which is incorporatedherein by reference in its entirety.

It is calculated by the percentage ratio of the fine particle mass(formerly fine particle dose) to the delivered dose.

The delivered dose is calculated from the cumulative deposition in theapparatus, while the fine particle mass is calculated from thedeposition of particles having a diameter <5.0 micron.

In the context of the present invention, the formulation is defined asextrafine formulation when, upon inhalation, the active ingredients aredelivered with a fraction of particles having a particle size equal toor lower than 2.0 micron equal to or higher than 20%.

With the term “mid FPF” is defined as the fraction of delivered dosehaving a particle size comprised between 0.2.0 and 5.0 micron.

The expression “physically stable in the device before use” refers to aformulation wherein the active particles do not substantially segregateand/or detach from the surface of the carrier particles both duringmanufacturing of the dry powder and in the delivery device before use.The tendency to segregate can be evaluated according to Staniforth etal. J. Pharm. Pharmacol. 34, 700-706, 1982, which is incorporated hereinby reference inits entirety, and it is considered acceptable if thedistribution of the active ingredient in the powder formulation afterthe test, expressed as relative standard deviation (RSD), does notchange significantly with respect to that of the formulation before thetest.

The expression “chemically stable” refers to a formulation that, uponstorage, meets the requirements of the EMEA Guideline CPMP/QWP/122/02,which is incorporated herein by reference in its entirety, referring to‘Stability Testing of Existing Active Substances and Related FinishedProducts’.

The term “surface coating” refers to the covering of the surface of thecarrier particles by forming a film of magnesium stearate around saidparticles. The thickness of the film has been estimated by X-rayphotoelectron spectroscopy (XPS) to be approximately of less than 10 nm.The percentage of surface coating indicates the extent by whichmagnesium stearate coats the surface of all the carrier particles.

The term “prevention” means an approach for reducing the risk of onsetof a disease.

The term “treatment” means an approach for obtaining beneficial ordesired results, including clinical results. Beneficial or desiredclinical results can include, but are not limited to, alleviation oramelioration of one or more symptoms or conditions, diminishment ofextent of disease, stabilized (i.e. not worsening) state of disease,preventing spread of disease, delay or slowing of disease progression,amelioration or palliation of the disease state, and remission (whetherpartial or total), whether detectable or undetectable. The term can alsomean prolonging survival as compared to expected survival if notreceiving treatment.

According to the Global Initiative for Asthma (GINA), which isincorporated herein by reference in its entirety, “uncontrolledpersistent asthma” is defined as a form characterized by daily symptoms,frequent exacerbations, frequent nocturnal asthma symptoms, limitationof physical activities, forced expiratory volume in one second (FEV₁)equal to or less than 80% predicted and with a variability higher than30%. According to the Global Initiative for Asthma (GINA) guidelines2014, which is incorporated herein by reference in its entirety,“partially uncontrolled asthma” is defined as a form characterized byless than twice a week daily symptoms, less than twice a month,nocturnal asthma symptoms, and a forced expiratory volume in one second(FEV₁) higher than 80% with a variability comprised between 20 and 30%.

According to the Global initiative for chronic Obstructive PulmonaryDisease (GOLD) guidelines which is incorporated herein by reference inits entirety, “severe COPD” is a form characterized by a ratio betweenFEV₁ and the Forced Vital Capacity (FVC) lower than 7 and FEV₁ between30% and 50% predicted. The very severe form is further characterized bychronic respiratory failure.

“Therapeutically effective dose” means the quantity of activeingredients administered at one time by inhalation upon actuation of theinhaler. Said dose may be delivered in one or more actuations,preferably one actuation (shot) of the inhaler. The term “actuation”refers to the release of active ingredients from the device by a singleactivation (e.g. mechanical or breath).

Wherein a numerical range is stated herein, the endpoints are included.

The present invention is directed to a process for the preparation of adry powder formulation for use in a dry powder inhaler (DPI) comprisinga carrier, and micronized particles of an anticholinergic, a long-actingβ₂-agonist (LABA), and, optionally, an inhaled corticosteroid (ICS), asactive ingredients.

The LABA active ingredient, that may be present in form ofpharmaceutically acceptable salts and/or solvate form thereof, can beselected from a group, which include, but it is not limited to,formoterol, salmeterol, indacaterol, olodaterol, vilanterol, and theultra-long-acting β2-adrenoreceptor agonist (uLABA) compound quoted withthe code AZD3199.

The anticholinergic, that is usually present in form of pharmaceuticallyacceptable inorganic salts, can be selected from a group which include,but it is not limited to, glycopyrronium bromide or chloride, tiotropiumbromide, umeclidinium bromide, aclidinium bromide, and the compoundquoted with the code GSK 233705.

The ICS, that may be anhydrous or present in form of hydrates, can beselected from a group which include, but it is not limited to,beclomethasone dipropionate and its monohydrate form, budesonide,fluticasone propionate, fluticasone furoate, and mometasone furoate.

Preferably, the LABA is formoterol fumarate dihydrate, the ICS isbeclometasone dipropionate, and the anticholinergic is glycopyrroniumbromide.

The carrier A) is constituted of a) a fraction of coarse excipientparticles and a fraction b) constituted of micronized excipientsparticles, and a salt of a fatty acid as an additive contributing toimprove the respirable fraction.

The coarse excipient particles consist of 80 to 95 percent by weight ofparticles of a physiologically acceptable excipient having a mass mediandiameter equal to or higher 175 micron.

Advantageously, all the coarse particles have a mass diameter in therange comprised between 100 and 600 micron.

In certain embodiments of the invention, the mass diameter of saidcoarse particles might be between 150 and 500 micron, preferably between200 and 400 micron.

In a preferred embodiment of the invention, the mass diameter of thecoarse particles is comprised between 210 and 360 micron.

In general, the skilled person shall select the most appropriate size ofthe coarse excipient particles if commercially available or by sieving,using a proper classifier.

Advantageously, the coarse excipient particles may have a relativelyhighly fissured surface, that is, on which there are clefts and valleysand other recessed regions, referred to herein collectively as fissures.The “relatively highly fissured” coarse particles can be defined interms of fissure index and/or rugosity coefficient as described in WO01/78695 and WO 01/78693, which are incorporated herein by reference intheir entireties, and they could be characterized according to thedescription therein reported. Advantageously, the fissure index of saidcoarse particles is of at least 1.25, preferably of at least 1.5, morepreferably of at least 2.0.

Said coarse particles may also be characterized in terms of tappeddensity or total intrusion volume measured as reported in WO 01/78695,which is incorporated herein by reference in its entirety.

The tapped density of said coarse particles could advantageously be lessthan 0.8 g/cm³, preferably 0.8 to 0.5 g/cm³. The total intrusion volumecould be of at least 0.8 cm³, preferably at least 0.9 cm³.

The fraction of micronized particles b) comprises of 19.6 to 4.9 percentby weight of particles of a physiologically acceptable excipient whereinat least 90% of said particles have a volume diameter lower than 15micron, preferably lower than 12 micron. Advantageously, the volumemedian diameter of said particles is 3 to 7 micron, preferably 4 to 6micron and no more than 10% of said particles have a diameter lower than2.5 micron, preferably lower than 2.0 micron.

Advantageously, the fine and coarse excipient particles may consist ofany pharmacologically inert, physiologically acceptable material orcombination thereof; preferred excipients are those made of crystallinesugars, in particular lactose; the most preferred are those made ofα-lactose monohydrate.

Preferably, the coarse excipient particles and the fine excipientparticles both consist of α-lactose monohydrate.

Said fraction b) further comprises 0.1 to 0.4 percent by weight of asalt of fatty acids such as lauric acid, palmitic acid, stearic acid,behenic acid, or derivatives (such as esters and salts) thereof.Specific examples of such salts of fatty acids are: magnesium stearate;sodium stearyl fumarate; sodium stearyl lactylate; sodium laurylsulphate, magnesium lauryl sulphate.

The preferred salt of fatty acid is magnesium stearate.

Advantageously, if it is used as the additive, magnesium stearate coatsthe surface of the coarse and micronized excipient particles a) and b)in such a way that the extent of the surface coating is at least of 5%more advantageously, higher than 10%.

The extent to which the magnesium stearate coats the surface of theexcipient particles may be determined by X-ray photoelectronspectroscopy (XPS), a well-known tool for determining the extent as wellas the uniformity of distribution of certain elements on the surface ofother substances. In the XPS instrument, photons of a specific energyare used to excite the electronic states of atoms below the surface ofthe sample, Electrons ejected from the surface are energy filtered via ahemispherical analyser (HSA) before the intensity for a defined energyis recorded by a detector. Since core level electrons in solid-stateatoms are quantized, the resulting energy spectra exhibit resonancepeaks characteristic of the electronic structure for atoms at the samplesurface.

Typically XPS measurements are taken on an Axis-Ultra instrumentavailable from Kratos Analytical (Manchester, UK) using monochromated AlKα radiation. (1486.6 eV) operated at 15 mA emission current and 10 kVanode potential (150 W). A low energy electron flood gun is used tocompensate for insulator charging. Survey scans, from whichquantification of the detected elements are obtained, are acquired withanalyser pass energy of 160 eV and a 1 eV step size. High-resolutionscans of the C 1s, O 1s, Mg 2s, N 1s and Cl 2p regions are acquired withpass energy of 40 eV and a 0.1 eV step size. The area examined isapproximately 700 μm×300 m for the survey scans and a 110 μm diameterspot for the high-resolution scans.

In the context of the present invention, it is possible to calculate byXPS both the extent of coating and the depth of the magnesium steratefilm around the lactose particles. The extent of magnesium stearate(MgSt) coating is estimated using the following equation:% MgSt coating=(% Mg_(sample)/% Mg_(ref))×100

where:

Mg_(sample) is the amount of Mg in the analyzed mixture; and

Mg_(ref) is the amount of Mg in the reference sample of commerciallyavailable MgSt.

Usually the values are calculated as a mean of two differentmeasurements. Typically, an accuracy of 10% is quoted for routinelyperformed XPS experiments.

Alternatively, when the excipient particles are made of lactose,preferably of alpha-lactose monohydrate, the extent of surface coatingmay be determined by water contact angle measurement, and then byapplying the equation known in the literature as Cassie and Baxter, forexample cited at page 338 of Colombo I et a II Farmaco 1984, 39(10),328-341, which is incorporated herein by reference in its entirety, andreported below.cos ϑ_(mixture) =f _(MgSt) cos ϑMgSt+f _(lactose) cos ϑ_(lactose)

where:

f_(MgSt) and f_(lactose) are the surface area fractions of magnesiumstearate and of lactose;

ϑ_(MgSt) is the water contact angle of magnesium stearate;

ϑ_(lactose) is the water contact angle of lactose; and

ϑ_(mixture) it are the experimental contact angle values.

For the purpose of the present invention, the contact angle may bedetermined with methods that are essentially based on a goniometricmeasurement. These imply the direct observation of the angle formedbetween the solid substrate and the liquid under testing. It istherefore quite simple to carry out, being the only limitation relatedto possible bias stemming from intra-operator variability. It should behowever underlined that this drawback can be overcome by adoption of afully automated procedure, such as a computer assisted image analysis, Aparticularly useful approach is the sessile or static drop method whichis typically carried out by depositing a liquid drop onto the surface ofthe powder in form of disc obtained by compaction (compressed powderdisc method).

Within the limits of the experimental error, a good consistency has beenfound between the values of extent of coating as determined by XPSmeasurements, and those as estimated by the theoretical calculationsbased on the Cassie and Baxter equation.

The extent to which the magnesium stearate coats the surface of theexcipient particles may also be determined by scanning electronmicroscopy (SEM), a well-known versatile analytical technique.

Such microscopy may be equipped with an EDX analyzer (an ElectronDispersive X-ray analyzer), that can produce an image selective tocertain types of atoms, for example magnesium atoms. In this manner itis possible to obtain a clear data set on the distribution of magnesiumstearate on the surface of the excipient particles.

SEM may alternatively be combined with IR or Raman spectroscopy fordetermining the extent of coating, according to known procedures.

More advantageously, the ratio among the fraction of coarse particlesa), the micronized excipient particles, and magnesium stearate shall becomprised between 85:14.7:0.3 and 90:9.8:0.2 by weight, preferably90:9.8:0.2 by weight.

Advantageously, the whole amount of coarse particles a) are mixed withthe whole amount of magnesium stearate and with a first portion of themicronized excipient particles.

Advantageously, said first portion is 40% to 60%, more advantageously 45to 55%, preferably 50%, based on the total weight of all micronizedexcipient particles.

The mixing may be performed in any suitable mixer, e.g. tumbler mixerssuch as Turbular™ for at least 5 minutes, preferably for at least 30minutes, more preferably for at least two hours.

In a general way, the skilled person shall adjust the time of mixing andthe speed of rotation of the mixer to obtain a homogenous mixture.

When spheronized coarse excipient particles are desired to obtainhard-pellets according to the definition reported above, the step ofmixing shall be typically carried out for at least four hours.

Since the mixing step does not alter the particle size, the personskilled in the art shall select the suitable size of the coarseexcipient particles, that of the micronized excipient particles as wellas that of magnesium stearate, either by sieving, by using a classifierto achieve the desired particle size distribution, being sure that finalparticle size of blend will correspond to the staring one.

Materials of the desired particle size distribution are alsocommercially available.

In one embodiment of the invention, the carrier A consisting of thecoarse excipient particles a), 50% of the micronized excipient particlesand the particles of magnesium stearate may be prepared by mixing in aTurbula™ mixer or in a dyna-MLX mixer at a rotation speed of 11 to 45rpm preferably 16 to 32 rpm, for a period of at least 30 minutes,preferably comprised between 60 and 300 minutes.

In step i), the carrier A), the micronized particles of the ICS, theLABA and the anti-muscarinic drug are poured in the vessel of a shakermixer having a wide and adjustable range of speed of rotation andinversion cycles.

It has indeed been found that said type of mixers is particularlysuitable due to their versatility. In fact, with said mixers, frequentchanges in the revolution cycles can be set in order to continuouslychange the powder flow inside the mixing and create powder flow patternswithin the drum and to increase mixing efficacy.

In a preferred embodiment of the invention, the dyna-MIX™ mixer isutilized.

The blend of step i) is mixed at a speed of rotation of at least 16r.p.m., preferably between 20 and 28 r.p.m, for a time of not less than60 minutes, preferably comprised between 60 and 120 minutes.

In step ii), the remaining part of the micronized physiologicallyacceptable excipient is added and mixed at a speed of rotation not lowerthan 16 rpm, preferably between 16 and 32 r.p.m., or a time of at least120 minutes, preferably between 120 and 180 minutes.

Contrary to what reported in the prior an it has indeed been found thatby adding micronized, and hence fine, particles of the excipient afterthe mixing of the active ingredients with the carrier, it is possible toreduce the de-aggregation of said active ingredients, and hence decreasethe respirable fraction.

Without being limited by the theory, this might be due that themicronized excipient particles cover the active ingredients particles,so partly preventing their de-aggregation.

Moreover, by proper controlling the amount of the micronized excipientparticles, it could be possible the extent of the reduction of therespirable fraction.

Optionally, the resulting mixture is sieved through a sieve. The skilledperson shall select the mesh size of the sieve depending on the particlesize of the coarse particles.

The blend of step ii) is finally mixed in any suitable mixer to achievean homogeneous distribution of the active ingredients.

The skilled person shall select the suitable mixer and adjust the timeof mixing and the speed of rotation of the mixer to obtain a homogenousmixture.

Advantageously, each active ingredient is present in the formulation ofthe invention in a crystalline form, more preferably with acrystallinity degree higher than 95%, even more preferably higher than98%, as determined according to known methods.

Since the powder formulation obtained with the process of the inventionshould be administered to the lungs by inhalation, at least 99% of saidparticles (d(v,0.99)) shall have a volume diameter equal to or lowerthan 10 micron, and substantially all the particles have a volumediameter comprised between 8 and 0.4 micron.

Advantageously, in order to better achieve the distal tract of therespiratory tree, 90% of the micronized particles of the ICS and LABAactive ingredients shall have a volume diameter lower than 6.0 micron,preferably equal to or lower than 5.0 micron, the volume median diametershall be comprised between 1.2 and 2.5 micron, preferably between 0.3and 2.2 micron, and no more than 10% of said shall have a diameter lowerthan 0.6 micron, preferably equal to or lower than 0.7 micron, morepreferably equal to or lower than 0.8 micron.

It follows that the width of the particle size distribution of theparticles of the ISC and LABA active ingredients, expressed as a span,shall be advantageously comprised between 1.0 and 4,0, moreadvantageously between 1.2 and 3.5 According the Chew et al J PharmPharmaceut Sci 2002, 5, 162-168, which is incorporated herein byreference in its entirety, the span corresponds to [d (v,0.9)−d(v,0.1)]/d(v,0.5).

In the case of the anticholinergic drug, in order to achieve both thedistal and upper tract of the respiratory tree, 90% of the micronizedparticles shall have a volume diameter equal to or lower than 8.0micron, preferably equal to or lower than 7.0 micron, the volume mediandiameter shall be comprised between 1.2 and 4.0 micron, preferablybetween 1.7 and 3.5 micron, and no more than 10% of said have a diameterlower than 0.5 micron, preferably equal to or lower than 0.6 micron,more preferably equal to or lower than 0.8 micron.

It follows that the width of the particle size distribution of theparticles of the anticholinergic drug, expressed as a span, shall beadvantageously comprised between 0.1.0 and 5.0, more advantageouslybetween 1.2 and 4.0.

The size of the particles of the active ingredients is determined bymeasuring the characteristic equivalent sphere diameter, known as volumediameter, by laser diffraction. In the reported examples, the volumediameter has been determined using a Malvern apparatus. However, otherequivalent apparatus may be used by the skilled person in the art.

In a preferred embodiment, the Helos Aspiros instrument (Sympatec GmbH,Clausthal-Zellerfeld, Germany) is utilized. Typical conditions are:Fraunhofer FREE or Fraunhofer HRLD algorithm, R1 (0.1/0.18-35 micron) orR2 (0.25/0.45-87.5 micron) lens, 1 bar pressure.

As for the particle size determination, a CV of ±30% for the d(v0,1) anda CV of ±20% for the d(v0,5), d(v0,9) and d(v0,99) are considered withinthe experimental error.

In a preferred embodiment of the invention, the LABA is formoterolfumarate dihydrate, the ICS is beclometasone dipropionate, theanticholinergic is glycopyrronium bromide, and the additive is magnesiumstearate.

Accordingly, in a particularly embodiment, the invention is directed toa powder formulation for inhalation for use in a dry powder inhaler,said powder comprising:

(A) a carrier, comprising:

(a) 80 to 95 percent by weight, based on the total weight of saidcarrier, of coarse particles of a physiologically acceptable excipienthaving a mean particle size of at least 175 μm; and

(b) 196 to 4.9 percent by weight, based on the total weight of saidcarrier, of micronized particles of a physiologically acceptableexcipient, and 0.1 to 0.4 percent by weight, based on the total weightof said carrier, of magnesium stearate; and

(B) micronized particles of glycopyrronium bromide, beclometasonedipropionate, and formoterol fumarate dihydrate, as active ingredients,

wherein said formulation is obtainable by a process comprising:

(i) mixing all of said coarse particles of a physiologically acceptableexcipient, all of said magnesium stearate, a first portion of saidmicronized particles of a physiologically acceptable excipient, all ofsaid micronized particles of glycopyrronium bromide, beclometasonedipropionate, and formoterol fumarate dihydrate in a vessel of a shakermixer at a speed of rotation not lower than 16 rpm for a time of notless than 60 minutes, to obtain a first mixture; and

(ii) adding the remaining part of said micronized particles of aphysiologically acceptable excipient to said first mixture, to obtain asecond mixture, and mixing said second mixture at a speed of rotationnot lower than 16 rpm for a time of at least 120 minutes;

whereby the extrafine particle fraction of each active ingredient iscomprised between 20 and 35%.

In a preferred embodiment, the extrafine particle fraction ofbeclometasone dipropionate, and formoterol fumarate dihydrate iscomprised between 20 and 35%, and the extrafine particle fraction ofglycopyrronium bromide is comprised between 20 and 30%.

Advantageously, in order to better achieve the distal tract of therespiratory tree, 90% of the micronized particles of beclometasonedipropionate (BDP), and formoterol fumarate dihydrate shall have avolume diameter lower than 6.0 micron, preferably equal to or lower than5.0 micron, the volume median diameter shall be comprised between 1.2and 2.5 micron, preferably between 1.3 and 2.2 micron, and no more than10% of said shall have a diameter lower than 0.6 micron, preferablyequal to or lower than 0.7 micron, more preferably equal to or lowerthan 0.8 micron.

It follows that the width of the particle size distribution of theparticles of the BDP and formoterol fumarate dihydrate, expressed as aspan, shall be advantageously comprised between 1.0 and 4.0, moreadvantageously between 1.2 and 3.5.

In the case of glycopyrronium bromide, in order to achieve both thedistal and upper tract of the respiratory tree, 90% of the micronizedparticles shall have a volume diameter equal to or lower than 8.0micron, preferably equal to or lower than 7.0 micron, the volume mediandiameter shall be comprised between 1,2 and 4.0 micron, preferablybetween 1.7 and 3.5 micron, and no more than 10% of said have a diameterlower than 0.5 micron, preferably equal to or lower than 0.8 micron,more preferably equal to or lower than 1.0 micron.

It follows that the width of the particle size distribution of theparticles of the anticholinergic drug, expressed as a span, shall beadvantageously comprised between 1.0 and 5.0, more advantageouslybetween 1.2 and 4.0.

More advantageously, it would also be preferable that the micronizedparticles of BDP have a Specific Surface Area comprised between 5.5 and7.0 m²/g, preferably between 5.9 and 6.8 m²/g, the micronized particlesof formoterol fumarate dihydrate have a Specific Surface Area comprisedbetween 5 and 7.5 mg, preferably between 5,2. and 6.5 m²/g, morepreferably between 5.5 and 5.8 m²/g, and the micronized particles ofglycopyrronium bromide have a Specific Surface Area comprised between1.8 and 5.0 m²/g preferably between 2.0 and 4.5 m²/g.

The Specific Surface Area is determined by Brunauer-Emmett-Teller (BET)nitrogen adsorption method according to a known procedure known.

All the micronized active ingredients utilized in the formulationaccording to the present invention may be prepared by processing in asuitable mill according to known methods.

In one embodiment of the present invention, they could be prepared bygrinding using a conventional fluid energy mill such as commerciallyavailable jet mill micronizers having grinding chambers of differentdiameters.

Depending on the type of the apparatus and size of the batch, the personskilled in the art shall suitably adjust the milling parameters such asthe operating pressure, the feeding rate and other operating conditionsto achieve the desired particle size. Preferably all the micronizedactive ingredients are obtained without using any additive during themicronization process.

In another embodiment of the present invention, the micronized particlesof glycopyrronium bromide may be prepared according to the processdisclosed in WO 2014/73987, which is incorporated herewith by referencein its entirety.

The powder formulation comprising micronized particles of glycopyrroniumbromide, beclometasone dipropionate, and formoterol fumarate dihydrateas active ingredients obtainable according to process of the presentinvention is physically and chemically stable, freely flowable andexhibits a good homogeneity of the active ingredients.

Moreover, the above powder formulation delivered through ahigh-performing DPI such as that disclosed in WO 2004/012801, which isincorporated herein by reference in its entirety, turned out totherapeutically equivalent to the corresponding pMDI formulation insolution.

The ratio between the carrier particles and the active ingredients willdepend on the type of inhaler used and the required dose.

The powder formulations of the present invention may be suitable fordelivering a therapeutic amount of all active ingredients in one or moreactuations (shots or puffs) of the inhaler.

Advantageously, the formulations of the present invention shall besuitable for delivering a therapeutically effective dose of all threeactive ingredients comprised between 50 and 600 g, preferably between100 and 500 μg.

For example, the formulations will be suitable for delivering 3-15 μg offormoterol (as fumarate dihydrate) per actuation, advantageously 5.5-6.5μg or 10-13 μg per actuation, preferably 6 or 0.12 μg per actuation;25-250 μg of beclometasone dipropionate (BDP) per actuation,advantageously 40-60 μg per actuation, or 80-120 μg per actuation, or160-240 μg per actuation; and 5-65 μg of glycopyrronium (as bromide),advantageously 5-15 μg per actuation or 20-30 μg per actuation,preferably 12.5 μg or 25 μg.

In a particular embodiment, the formulation is suitable for delivering 6μg of formoterol (as fumarate dihydrate) per actuation, 100 μg ofbeclometasone dipropionate, and 12.5 μg of glycopyrronium (as bromide)per actuation.

In another embodiment, the formulation is suitable for delivering 12 μgof formoterol (as fumarate dihydrate) per actuation, 200 g ofbeclometasone dipropionate, and 25 μg of glycopyrronium (as bromide) peractuation.

The dry powder formulation of the present invention may be utilized withany dry powder inhaler.

Dry powder inhaler (DPIs) can be divided into two basic types:

i) single dose inhalers, for the administration of single subdivideddoses of the active compound; each single dose is usually filled in acapsule; and

ii) multidose inhalers pre-loaded with quantities of active principlessufficient for longer treatment cycles.

The dry powder formulations of the present invention may be utilizedwith both multidose DPIs comprising a reservoir from which individualtherapeutic dosages can be withdrawn on demand through actuation of thedevice, or with single dose inhalers.

Typical multidose devices that may be used are, for instance, Diskus™ ofGlaxoSmithKhine, Turbohaler™ of AstraZeneca, Twisthaler™ of Schering,Clickhaler™ of Innovata, Spiromax™ of Teva, Novolizer™ of Meda, andGenuair™ of Almirall.

Examples of marketed single dose devices include Rotohaler™ ofGlaxoSmithKline, Handihaler™ of Boehringer Ingelheim, and Breezehaler™of Novartis.

Preferably, the powder formulation according to the invention is filledin a high-performing multidose DPI selected from the group consisting ofNEXTHaler™, and its variant disclosed in the application no.PCT/EP2015/063803, which is incorporated herewith by reference in itsentirety.

Other suitable high-performing multidose DPI are Novolizer™, andGenuair™.

To protect the DPIs from ingress of moisture into the formulation, itmay be desirable to overwrap the device in a flexible package capable ofresisting moisture ingress such as that disclosed in EP 1 760 008, whichis incorporated herein by reference in its entirety.

Administration of the formulation prepared according to the process ofthe present invention is indicated for the prevention and/or treatmentof chronic obstructive pulmonary disease (COPD) and asthma of all typesand severity.

The formulation prepared according to the process of the presentinvention is also indicated for the prevention and/or treatment offurther respiratory disorders characterized by obstruction of theperipheral airways as a result of inflammation and presence of mucussuch as chronic obstructive bronchiolitis.

In certain embodiments, said formulation is particularly suitable forthe prevention and/or treatment of severe and/or very severe forms COPD,and in particular for the maintenance treatment of COPD patients withsymptoms, airflow limitation and history of exacerbations.

Furthermore, it might be suitable for the prevention and/or treatment ofpersistent asthma and asthma in patients not controlled with medium orhigh doses of ICS in combination with LABAs.

Other features of the invention will become apparent in the course ofthe following descriptions of exemplary embodiments which are given forillustration of the invention and are not intended to be limitingthereof.

EXAMPLES Example 1. Preparation of the Carrier

Micronized alpha-lactose monohydrate (DFE Pharma, Germany) having thefollowing particle size: d(v0.1)=1.5 micron; d(v0.5)=3.6 micron; andd(v0.9)=7.5 micron was utilized.

About 1694 g of said micronized alpha-lactose monohydrate, about 69.2 gof magnesium stearate (Peter Greven, Germany) and about 31.13 kg offissured coarse particles of α-lactose monohydrate having a massdiameter of 212-355 micron (ratio 90:were fed into the vessel of aTurbula™ mixer (Willy A, Bachofen AG, Germany) and mixed. The mixing wascarried out for 240 minutes at a speed of rotation of 16 r.p.m.

Example 2. Preparation of the Dry Powder Formulation

Micronized formoterol fumarate dihydrate (FF) having the followingparticle size was used: d(v0.1)=0.9 micron; d(v0.5)=2.3 micron; andd(v0.9)=4.2 micron.

Beclometasone dipropionate (BDP) having the following particle size wasused: d(v0.1)=0.7 micron; d(v0.5)=1.5 micron; and d(v0.9)=2.8 micron.

Glycopyrronium bromide (GB) having the following particle size was used:d(v0.1)=0.4 micron; d(v0.5) 2.1 micron; d(v0.9)=5.5 micron.

The carrier as obtained in Example 1 was mixed in a dyna-MIX™ mixer withformoterol fumarate dihydrate, glycopyrronium bromide, and BDP at aspeed of rotation of 24 and 28 r.p.m. alternatively for the two rotationaxes for a time of 80 minutes.

Then 1694 g of micronised alpha-lactose monohydrate were added and mixedat a speed of rotation between 16 and 32 r.p.m, alternatively for thetwo rotation axes for a time of 150 minutes.

The resulting mixture was poured into a sieving machine available fromFrewitt (Fribourg, Switzerland) equipped with a 600 micron mesh sizesieve.

Upon sieving, the blend was finally mixed in a in the Dynamix mixer for60 minutes at a rotation speed of 24 and 32 r.p.m alternately to achievean homogeneous distribution of the active ingredients.

The ratio of the active ingredients to 10 mg of the carrier is 6 microg(μg) of FF dihydrate (theoretical delivered dose 4.5 μg), 100 microg(μg) of BDP and 12.5 microg (μg) of glycopyrronium bromide (theoreticaldelivered dose 10.0 μg).

The powder formulation was characterized in terms of the uniformity ofdistribution of the active ingredients and aerosol performances afterloading it in the multidose dry powder inhaler described in WO2004/012801, which is incorporated herein by reference in its entirety.

The uniformity of distribution of the active ingredients was evaluatedby withdrawing 10 samples from different parts of the blend andevaluated by HPLC

The results (mean value±RSD) are reported in Table 1.

The evaluation of the aerosol performance was carried out using the NextGeneration Impactor (NGI) according to the conditions reported in theEuropean Pharmacopeia 8.5th Ed 2015, par 2.9.18, pages 309-320, which isincorporated herein by reference in its entirety. After aerosolizationof 3 doses from the inhaler device, the NGI apparatus was disassembledand the amounts of drug deposited in the stages were recovered bywashing with a solvent mixture and then quantified by High-PerformanceLiquid Chromatography (HPLC).

The following parameters, were calculated: i) the delivered dose whichis the amount of drug delivered from the device recovered in the allparts of impactor; ii) the fine particle mass (FPM) which is the amountof delivered dose having a particle size equal to or lower than 5.0micron; iii) the extrafine FPM which is the amount of delivered dosehaving a particle size equal to or lower than 2.0 micron and/or equal toor lower than 1.0 micron and; iv) the mid FPM which is the amount ofdelivered dose having a particle size comprised between 2.0 and 5.0micron v) the fine particle fraction (FPF) which is the ratio betweenthe fine particle mass and the delivered dose: and vi) the MMAD.

The results (mean value±S.D) are reported in Table 1.

TABLE 1 Active ingredient FF Uniformity of distribution 100.5 (±1.5)Delivered Dose [μg] 5.1 Fine Particle Mass [μg] 2.9 Fine ParticleFraction [%] 54.8 Mid Fine Particle Mass μ] 1.24 Extrafine Particle Mass<2 μm [μg] 1.7 Extrafine Particle Mass <1 μm [μg] 0.6 Mid Fine particleFraction [%] 24.1 Extrafine Particle Fraction <2 μm [%] 32.5 ExtrafineParticle Fraction <1 μm [%] 11.7 MMAD [μm] 1.9 GB Uniformity ofdistribution 101.4 (±1.6) Delivered Dose [μg] 11.1 Fine Particle Mass[μg] 5.4 Fine Particle Fraction [%] 48.1 Mid Fine Particle Mass [μg] 2.4Extrafine Particle Mass <2 μm [μg] 2.9 Extrafine Particle Mass <1 μm[μg] 1.1 Mid Fine particle Fraction [%] 21.6 Extrafine Particle Fraction<2 μm [%] 26.4 Extrafine Particle Fraction <1 μm [%] 9.8 MMAD [μm] 1.9BDP Uniformity of distribution 100.5 (±1.8) Delivered Dose [μg] 88.5Fine Particle Mass [μg] 43.6 Fine Particle Fraction [%] 49.3 Mid FineParticle Mass [μg] 15.2 Extrafine Particle Mass <2 μm [μg] 28.5Extrafine Particle Mass <1 μm [μg] 12.4 Mid Fine particle Fraction [%]17.1 Extrafine Particle Fraction <2 μm [%] 32.1 Extrafine ParticleFraction <1 μm [%] 13.9 MMAD [μm] 1.6

Example 3. Reference Example from WO 2015004243

Two powder formulations according to the teaching of Example 1, 3, 4 and5 of WO 2015/004243, which is incorporated herein by reference in itsentirety were prepared.

Their aerosol performances, evaluated as reported in Example 2 of thepresent application, are reported in Table 2. MF is for mechano-fusionapparatus and CY is for Cyclomix™ apparatus.

TABLE 2 Batch Batch CY MF FF Delivered Dose [μg] 5.3 5.8 Fine ParticleMass [μg] 4.0 4.3 Fine Particle Fraction [%] 75.9 73.4 ExtrafineParticle Mass Fraction <2 μm [μg] 3.1 3.2 Mid Fine Particle Mass [μg]1.0 1.1 Extrafine Fine Particle Fraction <2 μm [%] 57.1 55.2 Mid FineParticle Fraction [%] 18.8 18.2 MMAD [μm] 1.1 1.2 GB Delivered Dose [μg]11.6 11.9 Fine Particle Mass [μg] 6.6 6.4 Fine Particle Fraction [%]57.2 53.8 Extrafine Particle Mass <2 μm [μg] 4.0 4.0 Mid Fine ParticleMass [μg] 2.6 2.5 Extrafine Particle Fraction <2 μm [%] 34.9 33.2 MidFine Particle Fraction [%] 22.3 20.6 MMAD [μm] 1.8 1.4 BDP DeliveredDose [μg] 90.6 95.7 Fine Particle Mass [μg] 64.5 66.9 Fine ParticleFraction [%] 71.2 69.9 Extrafine Particle Mass <2 μm [μg] 48.8 50.0 MidFine Particle Mass [μg] 15.7 16.9 Extrafine Particle Fraction <2 μm [%]53.9 52.3 Mid Fine Particle Fraction [%] 17.3 17.6 MMAD [μm] 1.1 1.1

Example 4, Reference Example from WO 201/076843

A pMDI HFA solution formulation according to the teaching of WO2011/076843 was prepared. Its aerosol performances, evaluated asreported in Example 2 of the present application, are reported in Table3.

TABLE 3 FF Delivered Dose [μg] 5.0 Fine Particle Mass [μg] 2.3 FineParticle Fraction [%] 45.7 Extrafine Particle Mass Fraction <2 μm [μg]2.0 Mid Fine Particle Mass [μg] 0.3 Extrafine Fine Particle Fraction <2μm [%] 45.7 Mid Fine Particle Fraction [%] 5.1 MMAD [μm] 1.0 GBDelivered Dose [μg] 10.6 Fine Particle Mass [μg] 4.8 Fine ParticleFraction [%] 45.1 Extrafine Particle Mass <2 μm [μg] 4.2 Mid FineParticle Mass [μg] 0.5 Extrafine Particle Fraction <2 μm [%] 40.2 MidFine Particle Fraction [%] 5.0 MMAD [μm] 1.0 BDP Delivered Dose [μg]87.8 Fine Particle Mass [μg] 40.4 Fine Particle Fraction [%] 46.0Extrafine Particle Mass <2 μm [μg] 35.9 Mid Fine Particle Mass [μg] 4.5Extrafine Particle Fraction <2 μm [%] 40.9 Mid Fine Particle Fraction[%] 5.2 MMAD [μm] 1.0

Example 5. Comparison of the FF/GB/BDP Dry Powder Formulation of theInvention with the Corresponding pMDI Solution Formulation of WO2011/076843

The study is designed to show that the 6/100/12.5 μg FF/GB/BDP drypowder formulation of the present invention delivered through the DPIdevice disclosed in WO 2004/012801, which is incorporated herein byreference, is therapeutically equivalent to the corresponding pMDI HFAsolution formulation of reference Example 4 in healthy volunteers. ThepMDI formulation is delivered with and without the Aerochamber Plus™Flow-Vu antistatic valved holding chamber.

Study Design:

Two parallel cohorts, open-label, randomized, 5-way crossover design.

Treatment: 8 single dose inhalations for a total dose of 48 microgramFF, 100 microgram GB, and 800 microgram BDP.

In order to obtain 20 evaluable subjects, approximately 25 healthyvolunteers will be randomized.

The study consists of 2 parallel subject cohorts, of five treatmentperiods each, with single-dose administration, separated by 16±2wash-out days between two consecutive treatment intakes.

Primary Objectives:

(1) To evaluate the total systemic exposure of 17-BMP (active metaboliteof BDP), FF, and GB as AUC₀₋₄ and C_(max); and

(2) To evaluate the lung availability of 17-BMP (active metabolite ofBDP), FF, and GB, assessed as systemic exposure (AUC₀₋₁ and C_(max))upon gastrointestinal charcoal blockage,

Secondary Objectives:

(1) To evaluate the pharamackinetic profile of BDP and additional PKparameters of 17-BMP, FF and GB assessed upon gastrointestinal charcoalblockage after administration: and

(2) To evaluate the general safety and tolerability profile with andwithout activated charcoal.

Endpoints:

Primary PK variables

17-BMP/FF/GB: AUC₁a Co.

Secondary PK variables

17-BMP/FF/GB: AUC_(0-∞), AUC_(0-30 min), t_(max) and t_(1/2).

BDP: AUC₀₋₁, C_(max) and t_(max)

Safety Variables

Adverse events and Adverse drug reactions.

Systolic Blood Pressure, Diastolic Blood Pressure, Heart Rate.

Measurements and Recording:

Pharmacokinetics Measures

-   -   BDP/17-BMP: 10 blood samples will be taken at the following time        points: pre-dose (within 60 min from dosing), 10, 15 and 30 min,        1, 2, 4, 8, 12 and 24 hours post-dose.    -   FF: 10 blood samples will be taken at the following time points:        pre-dose (within 60 min from dosing), 10, 15 and 30 min, 1, 2,        4, 8, 12 and 24 hours post-dose.    -   GB: 13 blood samples will be taken at the following time points:        pre-dose (within 60 min from dosing), 10, 15 and 30 min, 1, 2,        4, 8, 12, 24, 32, 48 and 72 hours post-dose.        Safety Measures    -   Blood Pressure and local safety ECG: recording will be done        -   At screening to evaluate subject inclusion.        -   At each Period to evaluate subject safety, at the following            time points: pre-dose (within 60 min from dosing), 10 min, 1            and 72h post-dose.    -   Clinical chemistry and serology; 1 blood sample will be taken at        screening (fasting condition from at least 10 hours).    -   Hematology: 1 blood sample will be taken at screening (fasting        condition from at least 11 hours).    -   Serum pregnancy test (only for women of childbearing potential)        will be done at screening to evaluate subject inclusion.    -   Urine testing; urine samples will be taken for urinalysis, drug        panel and cotinine test, at screening,    -   Urine pregnancy test (only for women of childbearing potential)        will be done at each Period to evaluate subject inclusion (at        randomization only) and subject safety        Statistical Methods:        Primary PK Variables    -   17-BMP, FF and GB C_(max) and AUC₀₋₄ (with/without activated        charcoal) will be log-transformed and analysed using a linear        model including treatment, sequence, period and subject within        sequence as fixed effects. For all the foreseen comparisons, the        ratios of adjusted geometric means will be calculated with their        90% two-sided confidence intervals (CIs).        Secondary PK Variables    -   17-BMP, FF and GB AUC₀₋₁, AUC_(0-30 min), and t_(1/2)        (with/without activated charcoal), BDP C_(max) and AUC₀₋₁        (with/without activated charcoal) will be log-transformed and        analysed using a linear model including treatment, sequence,        period and subject within sequence as fixed effects. For all the        foreseen comparisons, the ratios of adjusted geometric means        will be calculated with their 90% two-sided confidence intervals        (CIs).    -   17-BMP, BDP, FF and GB t_(max) (with/without activated charcoal)        will be analysed using the Wilcoxon signed rank test on        untransformed data and the Hodges-Lehmann nonparametric estimate        of location shift for all the foreseen comparisons.        Safety Variables

The number and the percentage of subjects who experience at least oneTEAE, drug-related TEAL serious TEAE, severe TEAE, TEAE leading to studydrug discontinuation and TEAE leading to death, as well as the number ofevents, will be summarized by treatment and overall.

The mean absolute value and the mean change from pre-dose (on the sameday in each treatment period) to each post-dose time-point in BloodPressure and Heart Rate will be calculated with their 95% CIs bytreatment.

Where a numerical limit or range is stated herein, the endpoints areincluded. Also, all values and subranges within a numerical limit orrange are specifically included as if explicitly written out.

As used herein the words “a” and “an” and the like carry the meaning of“one or more.”

Obviously, numerous modifications and variations of the presentinvention are possible in light of the above teachings. It is thereforeto be understood that, within the scope of the appended claims, theinvention may be practiced otherwise than as specifically describedherein.

All patents and other references mentioned above are incorporated infull herein by this reference, the same as if set forth at length.

The invention claimed is:
 1. A process for preparing a powderformulation for inhalation for use in a dry powder inhaler, said powdercomprising: (A) a carrier, comprising: (a) 80 to 95 percent by weight,based on the total weight of said carrier, of coarse particles of aphysiologically acceptable excipient having a mean particle size of atleast 175 μm; and (b) 19.6 to 4.9 percent by weight, based on the totalweight of said carrier, of micronized particles of a physiologicallyacceptable excipient, and 0.1 to 0.4 percent by weight, based on thetotal weight of said carrier, of a salt of a fatty acid; and (B)micronized particles of an anti-muscarinic drug, a long-actingβ₂-agonist, and optionally, an inhaled corticosteroid, as activeingredients, wherein said anti-muscarinic drug comprises glycopyrroniumbromide, and said long-acting β₂-agonist comprises formoterol fumaratedihydrate, said process comprising: (i) mixing all of said coarseparticles of a physiologically acceptable excipient; all of said salt ofa fatty acid; a first portion of said micronized particles of aphysiologically acceptable excipient; and all of said micronizedparticles of said long-acting β₂-agonist, said anti-muscarinic drug,and, optionally, said inhaled corticosteroid in a vessel of a shakermixer at a speed of rotation not lower than 16 rpm for a time of notless than 60 minutes, to obtain a first mixture; and (ii) adding theremaining part of said micronized particles of a physiologicallyacceptable excipient to said first mixture, to obtain a second mixture,and mixing said second mixture at a speed of rotation not lower than 16rpm for a time of at least 120 minutes, to obtain said formulation,wherein the extrafine particle fraction of each active ingredient isfrom 20 to 35%.
 2. The process according to claim 1, further comprising:(iii) further mixing said formulation obtained in (ii) to achieve ahomogeneous distribution of said active ingredients.
 3. The processaccording to claim 1, wherein said first portion of said micronizedparticles of a physiologically acceptable excipient is 40% to 60% byweight, based on the total weight of al of said micronized particles ofa physiologically acceptable excipient.
 4. The process according toclaim 1, wherein said long-acting β₂-agonist further comprises at leastone selected from the group consisting of formoterol, salmeterol,indacaterol, olodaterol, and vilanterol.
 5. The process according toclaim 1, wherein said anti-muscarinic drug further comprises at leastone selected from the group consisting of glycopyrronium chloride,tiotropium bromide, umeclidinium bromide, and aclidinium bromide.
 6. Theprocess according to claim 1, wherein said inhaled corticosteroid isselected from the group consisting of beclomethasone dipropionate,beclomethasone dipropionate monohydrate, budesonide, fluticasonepropionate, fluticasone furoate, and mometasone furoate.
 7. The processaccording to claim 1, wherein said inhaled corticosteroid isbeclometasone dipropionate.
 8. The process according to claim 1, whereinsaid salt of a fatty acid is selected from the group consisting ofmagnesium stearate, sodium stearyl fumarate, sodium stearyl lactylate,sodium lauryl sulfate, and magnesium lauryl sulfate.
 9. The processaccording to claim 8, wherein said salt of a fatty acid is magnesiumstearate.
 10. A powder formulation for inhalation for use in a drypowder inhaler, said powder comprising: (A) a carrier, comprising: (a)80 to 95 percent by weight, based on the total weight of said carrier,of coarse particles of a physiologically acceptable excipient having amean particle size of at least 175 μm; and (b) 19.6 to 4.9 percent byweight, based on the total weight of said carrier, of micronizedparticles of a physiologically acceptable excipient, and 0.1 to 0.4percent by weight, based on the total weight of said carrier, ofmagnesium stearate; and (B) micronized particles of glycopyrroniumbromide, beclometasone dipropionate, and formoterol fumarate dihydrate,as active ingredients, wherein said formulation is obtainable by aprocess comprising: (i) mixing all of said coarse particles of aphysiologically acceptable excipient; all of said magnesium stearate; afirst portion of said micronized particles of a physiologicallyacceptable excipient; and all of said micronized particles ofglycopyrronium bromide, beclometasone dipropionate, and formoterolfumarate dihydrate in a vessel of a shaker mixer at a speed of rotationnot lower than 16 rpm for a time of not less than 60 minutes, to obtaina first mixture; and (ii) adding the remaining part of said micronizedparticles of a physiologically acceptable excipient to said firstmixture, to obtain a second mixture, and mixing said second mixture at aspeed of rotation not lower than 16 for a time of at least 120 minutes,to obtain said formulation, wherein the extrafine particle fraction ofeach active ingredient is from 20 to 35%.
 11. The powder formulationaccording to claim 10, wherein said process further comprises: (iii)further mixing said formulation obtained in (ii) to achieve anhomogeneous distribution of the active ingredients.
 12. The powderformulation according to claim 10, wherein said first portion of saidmicronized particles of a physiologically acceptable excipient is 40% to60% by weight, based on the total weight of all of said micronizedparticles of a physiologically acceptable excipient.
 13. The powderformulation according to claim 10, whereby said extrafine particlefraction of beclometasone dipropionate, and formoterol fumaratedihydrate is from 20 to 35%, and said extrafine particle fraction ofglycopyrronium bromide is from 20 to 30%.
 14. The powder formulationaccording to claim 10, wherein said physiologically acceptable excipientis alpha-lactose monohydrate.
 15. The powder formulation according toclaim 10, wherein the coarse particles have a mass diameter of 210 to360 μm.
 16. A dry powder inhaler device, containing a dry powderformulation according to claim
 10. 17. A method for the treatment of aninflammatory and/or obstructive airways disease, comprisingadministering to a subject in need thereof an effective amount of a drypowder formulation according to claim
 10. 18. The method according toclaim 17, wherein said disease is asthma or chronic obstructivepulmonary disease (COPD).